Technology Development Unit

技术开发股：项目摘要 西北大学U54癌症染色质纳米成像中心(NU-CCNIC)的总体目标 是开发和部署多尺度染色质纳米成像平台，以及分子分析和 用于表征与癌症相关的染色质结构和转录模式的计算模型 干细胞(CSCs)与耐药表型。我们寻求在开发过程中填补三个技术空白 这样一个纳米成像平台。(1)多尺度挑战源于需要研究染色质是如何 在广泛的范围内调节基因表达，从DNA的直径(~2 nm)到 核小体(~10 nm)到染色质结构域(~100 nm)到细胞核(~10微米)，以及到一群细胞 解释了细胞间的异质性。(2)多路复用的分子成像挑战是由于 需要与3D染色质结构共同注册的关键基因和分子调节器的数量 数据。以及(3)时间动力学挑战需要在广泛的时间范围内工作的能力 研究可能在几分钟内发生的短期过程，如染色质重塑和长期 可能在几周内进展的过程，例如癌细胞出现化疗耐药。没有单程票。 技术满足了所有这三个要求。 我们应对这三个挑战的解决方案是将三个技术系列有机地关联起来： 染色质扫描电子显微镜(ChromSTEM)，光谱单分子定位 显微镜(SSMLM)和部分波谱(PWS)纳米传感显微镜。在注册这些之后 1纳米精度的技术，它们将共同满足多尺度、多路复用和动态 要求。我们将在技术方面实现三个目标。(1)开发下一代sSMLM，使其能够在 原理，无限多路传输，具有最高的光子利用率和1 nm的空间精度。(2)发展 相关ChromSTEM-sSMLM-PWS成像，实现1 nm精度的跨通道配准 由新的基于量子点的纳米组装受信标记辅助。以及(3)发展多尺度 阐明染色质结构与病因学关系的分子模拟平台 用纳米成像平台成像的CSCs中的转录重编程。建议的技术 开发是由研究试验台(RTB)阐明表观遗传学和染色质的需求驱动的 CSC过程中转录可塑性的驱动因素，特别是对 化疗，并探索将干细胞重新编程出干态作为治疗的可行性。 策略。RTB调查人员的持续反馈将与技术平台的优化相结合， 从长远来看，使我们的U54技术能够影响到更广泛的癌症研究社区 超越细胞癌生物学成像研究网络。

Technology Development Unit: PROJECT SUMMARY The overall goal of the U54 Northwestern University Center for Chromatin Nanoimaging in Cancer (NU-CCNIC) is to develop and deploy a multi-scale chromatin nanoimaging platform together with molecular analyses and computational modeling to characterize chromatin structure and transcriptional patterns associated with cancer stem cells (CSCs) and the chemoresistance phenotype. We seek to fill three technology gaps in developing such a nanoimaging platform. (1) The multi-scale challenge stems from the need to investigate how chromatin regulates gene expression across a wide range of scales, from the diameter of DNA (~2 nm) to the size of nucleosomes (~10 nm) to chromatin domains (~100 nm) to cell nuclei (~10 µm) and to a population of cells to account for intercellular heterogeneity. (2) The multiplexed molecular imaging challenge arises due to a large number of critical genes and molecular regulators that need to be co-registered with the 3D chromatin structural data. And (3) The temporal dynamics challenge requires the ability to work across a wide range of time scales to study short-term processes that may occur within minutes, such as chromatin remodeling, and long-term processes that may progress over weeks, such as the emergence of chemoresistance in cancer cells. No single technique satisfies all three requirements. Our solution to address these three challenges is to organically correlate three families of technologies: chromatin scanning transmission electron microscopy (ChromSTEM), spectroscopic single-molecule localization microscopy (sSMLM), and partial wave spectroscopic (PWS) nanosensing microscopy. After registering these technologies with 1 nm precision, they will collectively satisfy the multi-scale, multiplexing, and dynamics requirements. We will achieve three objectives in TECH. (1) Develop next-generation sSMLM that enables, in principle, unlimited multiplexing with the highest photon utilization and at 1 nm spatial precision. (2) Develop correlative ChromSTEM-sSMLM-PWS imaging that enables cross-modality registration at 1 nm precision assisted by novel quantum dot-based nano assembly fiduciary markers. And (3) Develop a multi-scale molecular modeling platform to elucidate the etiological relationship between chromatin structure and transcriptional reprogramming in CSCs imaged with the nanoimaging platform. The proposed technology development is driven by the needs of the Research Test-Bed unit (RTB) to elucidate epigenetic and chromatin drivers of transcriptional plasticity in CSC processes, in particular the development of adaptive resistance to chemotherapy, and to explore the feasibility of reprogramming CSCs out of the stem-states as a therapeutic strategy. Continuous feedback from RTB investigators will be integrated with the TECH platform's optimization, enabling, in the longer-term, our U54 technologies to impact the broader cancer research community within and beyond the Cellular Cancer Biology Imaging Research Network.